Bovine colostrum fraction as a serum substitute for the cultivation of mouse hybridomas

Summary Fractions of bovine colostrum were prepared and their ability to support the growth of mouse-mouse hybridomas in culture was tested. Whey was prepared from defatted colostrum by removal of casein using acid precipitation. An ultrafiltrate was obtained from cleared whey by filtration through membranes with a nominal molecular mass cut-off of 100 000 Da. Colostrum ultrafiltrate contained 1.16 g/l protein, 0.24 g/l immunoglobulin G (IgG) and less than 0.24 EU (endotoxin unit)/ml endotoxins. The effect of defatted colostrum, whey and ultrafiltrate as serum substitutes was examined by cultivation of hybridoma cells in minimal essential medium containing different concentrations of the supplements. Under optimal conditions in ultrafiltrate-supplemented medium, the maximal cell concentration was 35–40% of that obtained using 10% foetal bovine serum, and IgG production per cell was equal to that achieved using serum. In 1% defatted colostrum the maximum hybridoma concentration was about 30% of that in 10% serum, but at higher concentrations hybridoma growth was significantly reduced. The growth-promoting activity of whey was low. The results show that bovine colostrum ultrafiltrate provides a very attractive alternate to serum for production of monoclonal antibodies. Correspondence to: R. Pakkanen     

Comparison of selective media for coagulase-positive enterotoxigenic Staphylococcus aureus.

Different culture media were used to detect enterotoxigenic Staphylococcus aureus strains from peptone salt solution and inoculated minced meat. When recovery, reactions, and counting mistakes were considered, it was concluded that Baird-Parker agar was the best medium.     

Bovine αs1-casein gene sequences direct high level expression of human granulocyte-macrophage colony-stimulating factor in the milk of transgenic mice

The generation is reported of transgenic mice expressing human granulocyte-macrophage colony-stimulating factor (GM-CSF) or human erythropoietin (EPO) under the control of bovine s1- casein regulatory sequences. GM-CSF expression was specific to the mammary gland, and levels of human GM-CSF in transgenic mouse milk were in the range of mg ml–1. The specific activity of the milk GM-CSF was similar to that of the recombinant protein produced in Escherichia coli, and the glycosylation-derived size heterogeneity corresponded to that of the native human protein. In spite of the identical bovine regulatory sequences of the fusion genes, the levels of human EPO in transgenic mouse milk were 103--106 times lower than those of GM-CSF, ranging from 0.003 to 3 g ml–1. There appeared to be a positive correlation between the amount of EPO in the milk of lactating females and blood haematocrit values. In view of this, other type of constructs should be used to achieve more efficient EPO expression and to circumvent concomitantly-occurring adverse effects. In contrast, the high-level production of recombinant GM-CSF, its resemblance to the native mammalian protein, and mild adverse consequences of transgene expression imply that the current construct could be used for generation of larger GM-CSF transgenic anim als to produce this protein in quantities sufficient for therapeutic purposes     

Development of a Climate Choice meal concept for restaurants based on carbon footprinting

Purpose

Significant reductions in greenhouse gas emissions from food production and consumption can be made at the level of individual diet. Together with the food and beverage sector, consumers could play a significant role by making informed choices that benefit the environment and their own health. Communicating information on carbon footprints to consumers is challenging and should be made very simple, yet reliable. This sector is showing interest in using eco-design tools to decrease climate change impacts of their meals.

Methods

A long-term concept for communicating information on carbon footprints associated with meals was developed in Finland. The criteria for a Climate Choice meal were created through stakeholder dialogue, and three restaurant operators piloted the concept in 25 restaurants. In addition to climate change impacts, possibilities to include other sustainability criteria were reviewed. The concept was based on simplified carbon footprinting of raw material production and processing of ingredients for 105 commonly selected lunches. The carbon footprint calculations allowed the development of the Climate Choice meal concept, its criteria, and piloting the concept. Based on experiences from restaurants and consumers from the pilot phase, final criteria were developed.

Results and discussion

The Climate Choice meal concept was created using two alternative climate criteria: one for immediate implementation and another for future implementation, in cases where carbon footprinting is feasible for restaurants. The criteria for immediate implementation include a list of mainly plant-based ingredients with low carbon footprint. Regarding future criteria, it should be made easy enough for restaurants to estimate the carbon footprints of their meals, allowing labeling of meals when their carbon footprints are at least 25 % smaller than for an average meal. In addition to the two climate criteria, Climate Choice meals need to follow Finnish public catering nutritional recommendations, taking into account that fish species on the Red List of WWF’s Finnish seafood guide are prohibited.

Conclusions

To promote climate-friendly eating, a long-term concept rather than a short-term campaign is needed. There is interest among consumers and restaurants for information on food carbon footprints and sustainability. Lunch is regarded as a good opportunity for consumers to learn about climate-friendly eating. The main challenges are to produce sufficiently reliable background data and to raise consumer and the food and beverage sector interest and understanding of carbon footprints associated with food.

     

Type 2 diabetes whole-genome association study in four populations: the DiaGen consortium

Type 2 diabetes (T2D) is a common, polygenic chronic disease with high heritability. The purpose of this whole-genome association study was to discover novel T2D-associated genes. We genotyped 500 familial cases and 497 controls with >300,000 HapMap-derived tagging single-nucleotide-polymorphism (SNP) markers. When a stringent statistical correction for multiple testing was used, the only significant SNP was at TCF7L2, which has already been discovered and confirmed as a T2D-susceptibility gene. For a replication study, we selected 10 SNPs in six chromosomal regions with the strongest association (singly or as part of a haplotype) for retesting in an independent case-control set including 2,573 T2D cases and 2,776 controls. The most significant replicated result was found at the AHI1-LOC441171 gene region.     

Terrestrial organic matter quantity or decomposition state does not compensate for its poor nutritional quality for Daphnia

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Do all chlorophyll fluorescence emission wavelengths capture the spring recovery of photosynthesis in boreal evergreen foliage?

Chlorophyll a fluorescence (ChlF) is closely related to photosynthesis and can be measured remotely using multiple spectral features as solar‐induced fluorescence (SIF). In boreal regions, SIF shows particular promise as an indicator of photosynthesis, in part because of the limited variation of seasonal light absorption in these ecosystems. Seasonal spectral changes in ChlF could yield new information on processes such as sustained nonphotochemical quenching (NPQ_S) but also disrupt the relationship between SIF and photosynthesis. We followed ChlF and functional and biochemical properties of Pinus sylvestris needles during the photosynthetic spring recovery period to answer the following: (a) How ChlF spectra change over seasonal timescales? (b) How pigments, NPQ_S, and total photosynthetically active radiation (PAR) absorption drive changes of ChlF spectra? (c) Do all ChlF wavelengths track photosynthetic seasonality? We found seasonal ChlF variation in the red and far‐red wavelengths, which was strongly correlated with NPQ_S, carotenoid content, and photosynthesis (enhanced in the red), but not with PAR absorption. Furthermore, a rapid decrease in red/far‐red ChlF ratio occurred in response to a cold spell, potentially relating to the structural reorganization of the photosystems. We conclude that all current SIF retrieval features can track seasonal photosynthetic dynamics in boreal evergreens, but the full SIF spectra provides additional insight.     

Amino acid metabolism of experimental granulation tissue in vitro

1. The intracellular volume in granulation tissue was about 15% of the total urea space. 2. The experimental granuloma has a greater ability to retain amino acids during the proliferation phase than later during the synthesis of collagen. 3. The synthesis of collagen and other proteins by granulation tissue is related to the concentrations of proline and glutamic acid in the medium. 4. The rate of synthesis of proline from glutamic acid in granulation-tissue slices is greatest during collagen synthesis. It is enhanced by lactate. 5. Extracellular cations influence the synthesis of collagen and ouabain is inhibitory. Synthesis of other proteins is less sensitive in this respect. 6. It is suggested that the synthesis of collagen is related to the supply of certain amino acids, especially proline, and hence to the redox balance, and also to the function of the cell wall.     

Mechanisms for size-dependent protein segregation at immune synapses assessed with molecular rulers

Immunological synapses are specialized intercellular contacts formed by several types of immune cells in contact with target cells or antigen-presenting cells. A late-stage immune synapse is commonly a bulls-eye pattern of immune cell receptor-ligand pairs surrounded by integrin complexes. Based on crystal structures, the intermembrane distance would be ∼15 nm for many immune cell receptor-ligand pairs, but ∼40 nm for integrin-ligand pairs. Close proximity of these two classes of intermembrane bonds would require significant membrane bending and such proteins can segregate according to their size, which may be key for receptor triggering. However, tools available to evaluate the intermembrane organization of the synapse are limited. Here, we present what we believe to be a novel approach to test the importance of size in the intercellular organization of proteins, using live-cell microscopy of a size-series of fluorescently-labeled molecules and quantum dots to act as molecular rulers. Small particles readily colocalized at the synapse with MHC class I bound to its cognate natural killer cell receptor, whereas particles larger than 15 nm were increasingly segregated from this interaction. Combined with modeling of the partitioning of the particles by scaled-particle adsorption theory, these molecular rulers show how membrane-bending elasticity can drive size-dependent exclusion of proteins within immune synapses.     

Time scales of divergence and speciation among natural populations and subspecies of Arabidopsis lyrata (Brassicaceae)

? Premise of the study: Plant populations that face new environments adapt and diverge simultaneously, and both processes leave footprints in their genetic diversity. Arabidopsis lyrata is an excellent species for studying these processes. Pairs of populations and subspecies of A. lyrata represent different stages of divergence. These populations are also known to be locally adapted and display various stages of emerging reproductive isolation. ? Methods: We used nucleotide diversity data from 19 loci to estimate divergence times and levels of diversity among nine A. lyrata populations. Traditional distance-based methods and model-based clustering analysis were used to supplement pairwise coalescence-based analysis of divergence. ? Key results: Estimated divergence times varied from 130000 generations between North American and European subspecies to 39000 generations between central European and Scandinavian populations. In concordance with previous studies, the highest level of diversity was found in Central Europe and the lowest in North America and a diverged Russian Karhum?ki population. Local adaptation among Northern and central European populations has emerged during the last 39000 generations. Populations that are reproductively isolated by prezygotic mechanisms have been separated for a longer time period of ～70000 generations but still have shared nucleotide polymorphism. ? Conclusions: In A. lyrata, reproductively isolated populations started to diverge ～70000 generations ago and more closely related, locally adapted populations have been separate lineages for ～39000 generations. However, based on the posterior distribution of divergence times, the processes leading to reproductive isolation and local adaptation are likely to temporally coincide.